Puglisi Roberto, Severgnini Alex, Tava Aldo, Montedoro Marina
Istituto Sperimentale Italiano Lazzaro Spallanzani, Loc. La Quercia, 26027 Rivolta d'Adda (CR), Italy.
CREA-ZA Centro di Ricerca Zootecnia e Acquacoltura, Viale Piacenza 29, 26900 Lodi, Italy.
Food Technol Biotechnol. 2019 Mar;57(1):119-125. doi: 10.17113/ftb.57.01.19.5879.
Aqueous extracts were obtained at low temperature with the Naviglio technology from grapevine stalks (Merlot), marc (Merlot and Cabernet Sauvignon) and leaves (Merlot) as typical byproducts of winemaking industry, and their properties were evaluated cytofluorometrically on human dermal fibroblasts. Leaf extracts had the greatest total phenolic ((47.6±3.5) mg/g) and proanthocyanidin ((24.2±0.1) mg/g) contents compared to the others. The preliminary colorimetric MTT (3-[4,5-dimethylthiazole-2-yl]-2,5-diphenyltetrazolium bromide) assay individuated two consecutive non-toxic volume fractions of each extract (from 0.8 to 12.8%) that were adopted for three cytofluorometric tests. The first cell membrane test did not evidence any harmful effects against plasma membranes at the two non-toxic volume fractions. The second mitochondrial membrane test showed a decreased (p<0.01) percentage of cells ((15.7±8.3) (32.5±1.3) %) with active polarized mitochondrial membranes at the higher non-cytotoxic volume fractions of extracts from Cabernet Sauvignon marc in response to 4.5 mM HO, and from Merlot stalks (p<0.05) at 1.5 mM HO ((49.3±6.1) (64.6±2.4) %) and without HO ((89.7±2.4) (96.9±1.8) %), compared to the controls submitted to the same HO concentration. Conversely, mitochondrial activity of leaf extracts significantly (p<0.05) increased ((96.3±1.8) and (96.4±1.4) %) after treatment with 0.5 mM HO at both non-cytotoxic volume fractions compared to control ((88.2±1.1) %). Finally, as evidenced by the third oxidative status test, stalk extracts did not evidence relevant effects on the cellular oxidative state, while the extracts of marc and leaves demonstrated significantly medium (p<0.05) to highly (p<0.001) positive effects following exposure to HO ranging from 0.5 to 4.5 mM, compared to controls.
采用纳维廖技术在低温下从酿酒工业的典型副产品葡萄茎（梅洛）、葡萄皮渣（梅洛和赤霞珠）和葡萄叶（梅洛）中提取水提取物，并通过细胞荧光法对人皮肤成纤维细胞进行其性能评估。与其他提取物相比，叶提取物的总酚含量最高（(47.6±3.5)毫克/克），原花青素含量最高（(24.2±0.1)毫克/克）。初步的比色MTT（3-[4,5-二甲基噻唑-2-基]-2,5-二苯基四氮唑溴盐）试验确定了每种提取物的两个连续无毒体积分数（从0.8%到12.8%），并将其用于三项细胞荧光测试。第一次细胞膜测试未发现这两个无毒体积分数对质膜有任何有害影响。第二次线粒体膜测试显示，在赤霞珠葡萄皮渣提取物的较高非细胞毒性体积分数下，响应4.5 mM过氧化氢时，具有活性极化线粒体膜的细胞百分比降低（p<0.01）（(15.7±8.3)%对(32.5±1.3)%），在梅洛葡萄茎提取物中，在1.5 mM过氧化氢时（(49.3±6.1)%对(64.6±2.4)%）以及无过氧化氢时（(89.7±2.4)%对(96.9±1.8)%），与接受相同过氧化氢浓度处理的对照组相比。相反，在两个无毒体积分数下，用0.5 mM过氧化氢处理后，叶提取物的线粒体活性显著（p<0.05）增加（(96.3±1.8)%和(96.4±1.4)%），而对照组为(88.2±1.1)%。最后，如第三次氧化状态测试所示，茎提取物对细胞氧化状态没有显著影响，而葡萄皮渣和叶提取物在暴露于0.5至4.5 mM过氧化氢后，与对照组相比，显示出显著的中等（p<0.05）至高（p<0.001）的积极影响。
